Platforms for mAb Commercialization
-
Upload
kbi-biopharma -
Category
Health & Medicine
-
view
200 -
download
3
Transcript of Platforms for mAb Commercialization
Platforms for mAb commercialization
Abhinav A. Shukla, Ph.D.Senior Vice President
Development & ManufacturingKBI Biopharma, Durham NC
BPI West, San Francisco, February 27 – March 2, 2017
Durham, North Carolina- Cell Line Development- Cell Culture cGMP Manufacturing- Analytical QC, Formulation, Stability- Mass Spec Core Facility
RTP, North Carolina- Mammalian Process Development- Analytical development
Boulder, Colorado- Cell Line Development- Microbial Process Development- Microbial cGMP Manufacturing- Analytical, QC, Formulation, Stability- Particle Characterization Core Facility
The Woodlands, Texas- Cell Therapy
Manufacturing- Cell based assays
Contract Development & Manufacturing Organization
Programs in mammalian clinical manufacturing at KBI
• Primary emphasis 2011 – 2016 on clinical entry stage programs• ~ 10-14 IND filings per year supported via development &
manufacturing efforts• Also supported several stand-alone programs for process
characterization studies• Now emphasis shifting to include commercial launch process
development & manufacturing for mammalian cell culture programs• Boulder Colorado site already commercial ready for microbial
products
KBI Biopharma
Upstream Train I
Upstream Train II
ProA
VI
PolishVF
Bulk fill
KBI’s Cell Culture Manufacturing Facility
Purification Suite
2000L Prodn BRX200L Seed BRXWaveSF Harvest
2000L Prodn BRX200L Seed BRXWaveSF Harvest
3/9/20175 |
Mammalian cell culture expansion
• Leveraging significant capabilities in cell line development, analytical methods
• Development, formulation development & process development• Adding additional 2000L bioreactor in train II• Dedicated downstream purification for each train• Expanded buffer and media preparation integral with commercial suite• Increasing capacity to > 50 batches per year
0
5
10
15
20
25
30
2013‐14 2015 2016
New Products Mfg Batches
Cell Culture Manufacturing in KBI Biopharma (Durham, NC)
6
Biologics Commercialization
Pre-Clinical Phase I Phase II Phase III
Process DevelopmentProcess
CharacterizationProcess
ValidationProcess Monitoring
& Improvement
FIH Process• Deliver clinical process
quickly• Platform process• Clinical Supply
Submission & Approval
Lifecycle management
BLA Prep & PAI
Commercial Process• Deliver manufacturing process for
registrational trials and market• Design keeping large-scale manufacturing in
mind• Improve productivity, efficiency, robustness,
manufacturability, COGs• Analytical characterization and method
development
Process Characterization and Validation• Develop IPC strategy through understanding of process inputs and
outputs (design space)• Scale-down characterization and validation studies• Large-scale process validation to demonstrate process consistency• BLA preparation• Supporting documents for licensure inspections• Post-commercial process improvements (CI)• Post-commercial process monitoring
FIH process Commercial process
7Improvements in platform technology can enable one process development cycle
• Essential for biosimilars or highly accelerated programs
• Streamlined process characterization/validation effort
ProcessCharacterization
LifecycleManagementPlatform Application
IND BLA Commercial
Platform Technology Development
8
Outline• Where are mAb platforms today and do they enable a
single cycle of development?• FIH process• Commercial process
• Can a platform approach be taken for commercialization• Process characterization studies leading to definition of an in-
process control strategy (IPC) – how fast can these be completed?
• Scale-down validation studies• Conformance lots
• Commercialization in single-use manufacturing facilities
9
Investments in fundamental understanding of bioprocesses• Robust platforms can only be developed if there is a
strong understanding of the science of developing bioprocesses
• Multimodal chromatography• Platforms for non-mAbs (HIV vaccine proteins)• Improved Protein A resins
• Creates the ability to react quickly if an “unusual” observation is made
• All process decisions need to be made keeping large-scale production in mind
10
050
100150200250300350400450500
0.0% 20.0% 40.0% 60.0% 80.0% 100.0%
HCP
(ppm
)
Recovery
Capto MMC HCP Clearance25mM Tris pH 7.0 (baseline)
25mM Tris pH 7.0, 5% ethylene glycol
25mM Tris pH 7.0, 50mM arginine
25mM Tris pH 7.0, 50mM NaSCN
25mM Tris pH 7.0, 1M urea
25mM Tris pH 7.0, 1M ammonium sulfate
25mM Tris pH 7.0, 0.1M NaCl
25mM Tris pH 7.0, 0.5M ammonium sulfate
25mM Tris pH 7.0, 0.1M NaCl, 1M urea
Washes that disrupt protein-protein interactions
Conventional washes
log k’ = A – Blog(csalt) + C(csalt) k’ = (tr – tm )/tm
Wolfe, L., Barringer, C., Mostafa, S., Shukla, A. Multimodal chromatography: characterization of protein binding and selectivity enhancement through mobile phase modulators, Journal of Chromatography A, 1340, 151-156, 2014.
Multimodal chromatography
11
High capacity Protein A chromatography resins
Resin Vendor Matrix Ligand Modified Protein A domain
Mean particlesize (µm)
MabSelect SuReTM GE HealthcareHighly cross‐linked
agaroseAlkali‐stabilized
rProtein AB domain 85
MabSelect SuReTM LX GE HealthcareHighly cross‐linked
agaroseAlkali‐stabilized
rProtein A B domain 85
ToyopearlTM AF‐rProteinA HC‐650F† Tosoh Polymethacrylate
Alkali‐stabilized rProtein A
C domain 45
EshmunoTM A† EMD MilliporeCross‐linked
Polyvinyl EtherAlkali‐stabilized
rProtein A C domain 50
AmsphereTM A3† JSR Life Sciences Polymethacrylate
Alkali‐stabilized rProtein A
C domain 50
1 2 3 4 5 60
10
20
30
40
50
60
70
80mAb2
DB
C (g
/L)
Residence Time (min)
MabSelect SuRe MabSelect SuRe LX rProtein A HC-650F Eshmuno A Amsphere A3
mAb1 mAb2 mAb3 mAb40
1000
2000
3000
8000
9000
10000
HC
P Le
vel (
ppm
)
MabSelec SuRe MabSelect SuRe LX rProtein A HC-650F Eshmuno A Amsphere A3
CH505 Envelopes selected as vaccine immunogens
CH505 transmitted-founder (TF) and Env variants generated during viral evolution drove affinity maturation
of CH103 bnAb lineage
Antibody: UCAT/F gp120 Kd = ~200 nM
Env:
CH103
CH505 wk53
CH505 wk78
CH505 wk100
CH103 lineage intermediateantibodies
CH505 TF
CH505 wk136
12H.X. Liao et al. Nature 496: 469; 2013
• Parameters shaded in gray are defined across molecules. Parameters shaded in yellow require molecule specific optimization
• For all Env molecules the operating parameters, basal medium, feed type and some of the supplement additions have defined
• The need for additional supplements is molecule specific
• Reasons for supplement addition:
» Biocompatability in SU bioreactors
» Increase in productivity
ScaleTemperature Set point 37.0 ± 0.5°C Temperature Shift 33.0 ± 0.5°C on Day 6DO Set point 30%pH Set point 6.90 ± 0.1
Agitation (1-impeller) 50 rpm → 55 rpm
Air overlay 1.6 SLPMAir Sparge 0.5 SLPM Max. Oxygen Sparge 5 SLPMMax. CO2 Sparge 5 SLPM
Medium CD OptiCHO + 8 mM Glutamine
Target VCD 0.50 x 106 cells/mLBase 1M Sodium carbonate
Feed Type: LTI Feed A+B (1:1) 15% on Day 0, 10% current wv each on Days 3, 6, and 9
Supplement 1 addition: HT Supplement 1X current wv each on Days 0 and 4
Supplement 2 addition: CystineSupplement 3 addition: TyrosineSupplement 4 addition: Soy:Yeastolate Hydrolysate (2:3) 5g/L current wv each on Days 4 and 8
Supplement 5 addition: C1615Harvest Add 10g/L Hydrolysate on harvest
• Parameters shaded in gray are defined across molecules. Parameters shaded in yellow require molecule specific optimization
• Load and elution conditions for three of the unit operations require molecule specific definition given the heterogeneity of this class of molecules
• Env antigens structurally sensitive to hydrophobic surfaces, hence HIC not employed
1
10
100
1000
10000
100000
Log HCP
(pp
m)
Downstream Process
Platform HCP Clearance
TF Demo
TF ENG
TF GMP
w100 Demo
w100 ENG
w100 GMP
w78 Demo
w78 GMP
SEC‐HPLC % Main Peak
Sample ID TF Demo
TF ENG
TF GMP
w100 Demo
w100 ENG
w100GMP
w78Demo
w78 GMP
BDS 99.3% 98.9% 98.8% 98.9% 99.3% 99.2% 99.5% 99.6%
16
MAB PLATFORMS
17
Next generation mAb platforms• Driver
• High cell culture productivity is increasing interest in ultra-high loading polishing steps (> 100 mg/mL loading)
Genentech Biogen Millipore proposalProtein A
Viral Inactivation
Cation-exchange chromatography
Anion-exchange chromatography
Viral Filtration
UF/DF
Protein A
Viral Inactivation
AEX flowthrough
No salt Hydrophobic Interaction
Chromatography flowthrough
Viral Filtration
UF/DF
Protein A
Viral Inactivation
Anion-exchange flowthrough
Overloaded cation-exchange
chromatography
Viral Filtration
UF/DF
18
Which platform should I use?
19
Next generation mAb platforms• Platform processes for mAbs have hugely facilitated
the growth of mAbs as therapeutic agents• Rapid clinical entry with lower cost & resource burden
& significant time savings (gene to IND in ~ 12-14 months)
DS ProcessPlatform
DS ProcessPlatform
Cell line diversityCell line diversity
Media/feed type
diversity
Media/feed type
diversity
HCP level variabilityHCP level variability
Cell density variability
Cell density variability
HMW level variability
HMW level variability
Protein A
Viral Inactivation
AEX Based Polishing (Flow Through mode)
CEX Based Polishing
Viral Filtration
UF/DF
20Multimodal chromatography in next generation mAb platforms
• Mixed-mode has the simultaneous ability to clear HMW and HCP leading to mAb platforms with wider coverage
• Added advantage of ability to operate over wider conductivity range for loading
93.0
94.0
95.0
96.0
97.0
98.0
99.0
100.0
0.0 10.0 20.0 30.0 40.0 50.0 60.0 70.0 80.0 90.0 100.0
Capto S ImpAct pH 5.0
Eshmuno CPX pH 5.0
Fractogel SO3 pH 5.0
Capto MMC pH 7.5
Selectivity Curves for HMW Clearance
Mai
n Pe
ak (%
)
Increase selectivity
Accumulated Yield (%)
Hydrophobicity scale: Capto S < Fractogel SO3 < POROS HS50 < Nuvia cPrime < Capto MMC
21Success of mAb platforms that include multimodal chromatography
• Can successfully accommodate wide range of cell lines and cell culture feed streams into a single downstream platform
• Cell lines from KBI, Bioceros, Selexis, Cellca, Excellgene, Antitope, Life Technologies
0.0%
1.0%
2.0%
3.0%
4.0%
5.0%
6.0%
7.0%
8.0%
ProA AEX CEX BDS
%H
MW
mAb Platform HMW Clearance
mAb A
mAb B
mAb C
mAb D
mAb E
mAb F
mAb G
mAb H
mAb I0
5000
10000
15000
20000
25000
ProA AEX CEX BDS
rHC
P (p
pm)
mAb Platform rHCPClearance
mAb A
mAb B
mAb C
mAb D
mAb E
mAb F
mAb G
mAb H
mAb I
22
MAPPING PROCESS DESIGN SPACE (PROCESS CHARACTERIZATION AND SCALE-DOWN VALIDATION)
23
Quality by Design (QbD)• “Quality by design means designing and developing
manufacturing processes during the product development stage to consistently ensure a predefined quality at the end of the manufacturing process.” ICH Q10, FDA 2006
Process Design(Process Development)
Process ControlStrategyDefinition
ProcessValidation
Continued ProcessVerification
24
Process Design Space
Higher level of assurance of product quality
Manufacturing Efficiency and Flexibility Continuous process
improvement while maintaining product quality
Characterization Space
Design space
Control space
Design Space (ICH Q8, 2006): The multidimensionalcombination and interaction of input variables (e.g., material attributes) and process parameters that have been demonstrated to provide assurance of quality.
Need a high throughputscale-down model forthe process
25
Accelerating the Entire Product Development Lifecycle
26
Accelerating process characterization & scale-down validation studies• Small-scale bioreactors (1-10L working volume) have
been the traditional scale-down model in industry till date
• Accelerating PC/PV studies requires a high-throughput scale-down model
• Ambr250 as a scale-down model for cell culture processes
27
Mimicking “Cellular Environment” in SDMs.
Given the large sets of variables in a cell culture process, establishing a costand time-efficient SDM, mimicking a cellular environment similar to large scaleproduction bioreactor, is critical for conducting successful PC studies.
28
Matching key process indicators in SDMs
Comparison of time courses for viable cell growth and lactate profiles for two recombinant CHO cell lines inambr™ SDMs for a mAb and a Biosimilar. Matching cell growth and lactate profiles for CHO cell linesproducing a mAb and Biosimilar respectively were key process indicators and in turn dictated the processyield and product quality.
29
Comparison of SDMs across scales
30
Accelerated Upstream PC Timelines with high-throughput SDMsMonth 1.5
SDMQ USP
Month 5.5
N-1/N-2 Screening(40 x 3L Seed)
Harvest PC Work12 -15 Harvest conditions
Month 0
Raw Materials and Worst Case (20 x 3L and 1 round of ambr250 runs: 24 vessels)
Main Stage PC (3 rounds of ambr250 runs: 72 vessels)
Inoculum Studies(100 Shake Flasks and 4 Wave Runs)
Worst-case Linkage USP/DSP
Month 7.0
31
Scale-Down Process Validation Studies
• Scale-down validation studies in addition to large-scale process validation (conformance lots)
• Probe extremes in the process and demonstrate them to be acceptable
• Examples• Reprocessing validation – combine hold times with process
conditions that create the greatest stress on the protein• Intermediate hold times – combine hold times and
demonstrate releasable drug substance• Viral clearance studies• Impurity clearance studies
32
Conclusions• Robust scientific understanding is a pre-requisite for
developing robust platforms that can make single cycle development possible
• Highly generic and manufacturing friendly mAb platforms have been designed (gene to IND in 12-14 months)
• Process characterization & scale-down validation studies can be accelerated (6-9 months) by using high throughput cell culture platforms as the scale-down model
• Single-use manufacturing is now a viable commercial launch platform (long term may combine single use manufacturing & higher productivity continuous manufacturing)
33
mAb platforms
SU manufacturing
High through process development tools
Rapid process characterization & validation tools & approaches
Henry David Thoreau1817-1862, writer
Walden Pond Mahatma Gandhi1869 - 1948
34